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Raeusch. and Allophylus Serratus (Roxb.) Kurz Original research article GC-MS Analysis of Young Leaves of Allophylus cobbe (L.) Raeusch. and Allophylus serratus (Roxb.) Kurz. Chavan Reshama Bharat and Gaikwad Dattatraya Krishna Department of Botany, Shivaji University, Vidyanagar, Kolhapur, 416004, (MS) INDIA. ABSTRACT Background: Allophylus cobbe(L.) Raeusch. and Allophylus serratus (Roxb.)Kurz. (Sapindaceae) are medicinal plants traditionally used to cure bone fractures and other ailments in India. Objective: Present work was aimed at the identification of phytochemical compounds from young leaves of A. cobbe and A. serratus by applying gas chromatography mass spectrometry. Method: GC-MS technique was used for analysis of compounds. Results: It indicates 11 compounds from A. serratus and seven compounds from A. cobbe having mixture of terpenoids, alkanes and fatty acids. The major compounds found in A. cobbe were 1,1-diethoxy ethane (82.97%) (RT: 3.192min), phytol (7.07%) (RT: 43.690 min) and hexanoic acid (3.67%) (RT: 29.576 min) while in A. serratus 3-methyl butanol (53.16%) (RT: 3.223 min), 2-propenoic acid, 2-(dimethylamino) ethyl ester (27.08%) (RT: 13.94 min) and diisocytyl phthalate (4.49%) (RT: 52.584 min) were found in higher quantity. Phytol was detected from both species. Phytol possesses anti-radical, anti-cancer, antibacterial and anti-inflammatory properties as well as used in artificial synthesis of vitamin E and vitamin K in cosmetics and fragrance industry. Conclusion: The abundant presence of phytol in leaves of Allophylus species (7.07% in A. cobbe and 2.60% in A. serratus) suggests that these species may prove helpful in prevention of degenerative diseases and application of these herbs in cosmetics which may prove a cost effective natural alternative to current synthetic cosmetics. Presence of industrially important chemicals like phytol, 1,1-diethoxy ethane, hexanoic acid, diisocytyl phthalate, 2-propeonic acid, 2-(dimethylamino) ethyl ester underlines industrial importance of these species which needs further research on this aspect to focus in detail. Key words: GC-MS Analysis, Allophylus Cobbe, Allophylus Serratus, Young Leaves, phytol, medicinal plants. INTRODUCTION Submission Date: 08-12-2016; Revision Date: 24-01-2017; 5 6 6 Allophylus (family Sapindaceae) having about bone fractures, rashes, and stomach ache. Accepted Date: 18-03-2017 255 species found worldwide out of which Benzylamide is isolated from leaves of DOI: 10.5530/ijper.51.3.75 1 7 nine species are found in India. A. cobbe and A. cobbe (L.) Raeusch . A. serratus is used Correspondence: A. serratus are species found in Maharashtra against bone fractures.8 A. serratus (Kurz) Dr. Chavan Reshama Bharat, (M.Sc. Ph.D. Botany) 2 9 state of India. These two species are found leaves contain β-sitostrerol and phenac- Department of Botany, in various localities of Kolhapur district of etamide, an anti-ulcer phytochemical.10 Shivaji University, Kolhapur 3 416 004, INDIA. Maharashtra State. Most of the local people Kumar et al. observed osteogenic activity Phone no: 8722793734 in Maharashtra call both the species by a of rutin isolated from stem of A. serratus E mail: [email protected] same name- Tipan (in Marathi language).4 in in vitro studies.11 Chavan and Gaikwad The fresh leaves of these species are tied reported antibacterial activity of A. serratus over the bone fractures for a faster recovery.4 and A. cobbe.12 The fresh leaves as well as dry powder of In recent years, GCMS is being a helpful leaves are also taken with small amount tool to identify the phytochemicals from of jaggery at early morning with empty the medicinal plants and various natu- stomach, for relief from joint pains.4 Leaves ral products; because it is a non-destruc- of A. cobbe are used by local people against tive, direct and fast analytical method www.ijper.org 472 Indian Journal of Pharmaceutical Education and Research | Vol 51 | Issue 3 | Jul-Sep, 2017 Chavan RB and Gaikwad DK: GC-MS analysis of young leaves of Allophylus cobbe (L.) Raeusch. and Allophylus serratus (Roxb.) Kurz. for identification of terpenoids, fatty acids and other 62,000 patterns (computer software version 1.10 beta, phytochemicals as well as a very less plant material Shimadzu). is required. The phytochemical information of young leaves of RESULTS AND DISCUSSION these species is scanty. Hence, present investigation was The GCMS data of A. cobbe is represented in Table 1 and aimed at characterizing chemical components of these plate 1. It indicates that it is the mixture of terpenoids, species. alkanes and fatty acids. A total of seven compounds were identified with the help of their retention indices MATERIAL AND METHODS as well as by mass spectrometric data. The sample of Collection of plant material young leaves of A. cobbe comprised of 1,1-diethoxy Young leaves of A. cobbe and A. serratus were collected ethane (82.97%), mesostilbene glycol (2.57%), 1,1-diethoxy from forest of Panhala (Kolhapur district, Maharashtra propane (0.6%), hexanoic acid (3.67%), amyl nitrite (1.13%), eicosanoic acid, ethyl ester (2%) and phytol State, India). The material was washed with water, blotted (7.07%). Major phytochemicals detected were 1,1-diethoxy to dry and kept in oven at 60° C until gaining the ethane (retained by 3.192 min.) and phytol (retained by constant weight. The completely dried material was 43.690 min. from column). taken out, finely ground with Willey’s grinding machine and these powders were kept in airtight containers until From young leaves of A. serratus, 11 compounds use. have been detected as indicated in Table 2 and chro- matogram in plate 2. It is the mixture of terpenoids Extraction of Plant Material and fatty acids. Eleven co mpounds were identified with the help of NIST library and available data. It 5 g of dry powders of young leaves of A. serratus and comprises 3methyl -1-butanol (53.16%), 2-propenoic A. cobbe were taken and extracted at 60 °C in absolute acid,2-(dimethylamino) ethyl ester (27.08%), diisocy- ethanol (Merck) up to 8 hr by using soxhlet apparatus. tyl phthalate (4.49%), palmitic acid ethyl aster (3.51%), After completion of extraction, extract was taken out, 1,1,3-triethoxy propane (3.17%), phytol (2.6%), poured in evaporating dish, kept on water bath (60ºC) n-hexadecanoic acid (1.68%), Nonanoic acid,9-(3-ex- and evaporated to dryness. Resulting residue remained anylidenecyclo propylidene)-2-hydroxy methyl ethyl at bottom of evaporating dish was dissolved in 5 ml ester (1.17%), octadecanoic acid, ethyl ester (1.15%), absolute ethanol and used for GC-MS analysis. 1,1-diethoxy-3-methyl-butane (1.01%) and cyclopentyl 2-n-hexadecylcyclopentane (0.99%). Major compounds Chromatographic analysis found were 3-methyl 1-butanol (retention time-3.223 GC-MS analysis was performed on a Shimadzu (Tokyo, min) and propenoic acid, 2-(di methylamino) ethyl ester Japan) Make QP-2010 with nonpolar 60 M RTX 5MS (retention time-13.942 min). capillary column, full scan mode, injector mode-split, Previous studies by Priya et. al. indicated presence of 22 (split ratio 1:20), quadra pole mass selective detector different compounds in ethanolic extracts of leaves of (MSD), injection temperature 220ºC, GC-MS interface A. serratus, identified by GC-MS technique, in which temperature 230ºC, the injection volume was 1μl. cycloheptasiloxane, diethyl phthalate and hexasiloxane Helium was employed as carrier gas, at a pressure of 60 were major compounds.13 Present study gives different KPa; flow rate was 1ml/min. Mass spectra were detected result from that of the study by Priya et. al. which may at 70eV. Temperature programming was set as follows: be due to the difference between type of plant material column temperature was started from 60ºC (held for (in present analysis, only young leaves have been used), 2 min) and linearly increased by 5ºC/min to 130ºC (held the temperature programming and solvent extraction for 2min); after that it was increased by 4ºC/min to procedure. Various phytochemicals have been isolated 200ºC (held for 2 min); further it was increased by 8ºC/ from different parts of Allophylus species (See appendix). min. to 250ºC (held for 10 min). Total GC running time An anti-ulcer compound phenacetamide have been was 55 min. detected from leaves of A. serratus.10 Extraction of A. serratus leaves followed by column chromatography Identification of components over silica gel revealed the presence of various com- Interpretation of mass spectra of GC-MS was con- pounds like- pinitol, quercetin, rutin, luteolin-7-O-β- ducted using the database of National Institute of D-glucopyranoside, and apigenin-4’-O-β-D-gluco-side Standard and Technology (NIST) having more than which were tested for osteogenic activity.11 In vitro Indian Journal of Pharmaceutical Education and Research | Vol 51 | Issue 3 | Jul-Sep, 2017 473 Chavan RB and Gaikwad DK: GC-MS analysis of young leaves of Allophylus cobbe (L.) Raeusch. and Allophylus serratus (Roxb.) Kurz. osteogenic activity was studied using calvarial osteoblasts extract was tested by agar well diffusion method against obtained from Sprague Dawley rats by applying method bacteria-Staphyllococcus aureus and Bacillus subtilis.12 As of Ishizuya et al.11-14 Cultured osteoblasts at ~80% compared to cefotaxime, both aqueous and ethanolic confluence were trypsinised and treated with isolated extracts of young and mature leaves of AC and AS indi- compounds from plant extracts which indicated increase cated remarkable antibacterial activity against B. subtilis.12 in alkaline phosphatase activity due to rutin.11 Chavan A. cobbe exhibited significant nematicidal activity against and Gaikwad detected antibacterial activity of aqueous the root knot nematode Meloidogynae incognita.15 and ethanolic extracts of young and mature leaves of There are some reports on phytochemical studies of A.
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